Improved multiparametric scrape loading-dye transfer assay for
a simultaneous high-throughput analysis of gap junctional
intercellular communication, cell density and viability

J 2020

Improved multiparametric scrape loading-dye transfer assay for a simultaneous high-throughput analysis of gap junctional intercellular communication, cell density and viability

DYDOWICZOVÁ, Aneta; Ondřej BRÓZMAN; Pavel BABICA a Iva SOVADINOVÁ

Základní údaje

Originální název

Improved multiparametric scrape loading-dye transfer assay for a simultaneous high-throughput analysis of gap junctional intercellular communication, cell density and viability

Autoři

DYDOWICZOVÁ, Aneta; Ondřej BRÓZMAN; Pavel BABICA a Iva SOVADINOVÁ

Vydání

Scientific reports, LONDON, NATURE PUBLISHING GROUP, 2020, 2045-2322

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10500 1.5. Earth and related environmental sciences

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.380

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/20:00114682

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

LIVER EPITHELIAL-CELLS; WB-F344 RAT-LIVER; FUNCTIONAL EXPRESSION; MOLECULAR PERMEABILITY; HA-RAS; CONNEXIN; INHIBITION; CHANNELS; CX43; IMAGE

Štítky

143180, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 3. 6. 2025 20:32, Mgr. Michaela Hylsová, Ph.D.

Anotace

V originále

Gap junctional intercellular communication (GJIC) is a vital cellular process required for maintenance of tissue homeostasis. In vitro assessment of GJIC represents valuable phenotypic endpoint that could be effectively utilized as an integral component in modern toxicity testing, drug screening or biomedical in vitro research. However, currently available methods for quantifying GJIC with higher-throughputs typically require specialized equipment, proprietary software and/or genetically engineered cell models. To overcome these limitations, we present here an innovative adaptation of traditional, fluorescence microscopy-based scrape loading-dye transfer (SL-DT) assay, which has been optimized to simultaneously evaluate GJIC, cell density and viability. This multiparametric method was demonstrated to be suitable for various multiwell microplate formats, which facilitates an automatized image acquisition. The assay workflow is further assisted by an open source-based software tools for batch image processing, analysis and evaluation of GJIC, cell density and viability. Our results suggest that this approach provides a simple, fast, versatile and cost effective way for in vitro high-throughput assessment of GJIC and other related phenotypic cellular events, which could be included into in vitro screening and assessment of pharmacologically and toxicologically relevant compounds.

Návaznosti

EF16_013/0001761, projekt VaV

Název: RECETOX RI

GJ16-10775Y, projekt VaV

Název: Mezibuněčná komunikace zprostředkovaná mezerovými spoji jako cíl endokrinních disruptorů v testikulárních buňkách

Investor: Grantová agentura ČR, Mezibuněčná komunikace zprostředkovaná mezerovými spoji jako cíl endokrinních disruptorů v testikulárních buňkách

90051, velká výzkumná infrastruktura

Název: RECETOX II

Citovat

DYDOWICZOVÁ, Aneta; Ondřej BRÓZMAN; Pavel BABICA a Iva SOVADINOVÁ. Improved multiparametric scrape loading-dye transfer assay for a simultaneous high-throughput analysis of gap junctional intercellular communication, cell density and viability. Scientific reports. LONDON: NATURE PUBLISHING GROUP, 2020, roč. 10, č. 1, s. 1-17. ISSN 2045-2322. Dostupné z: https://doi.org/10.1038/s41598-020-57536-3.

@article{1729212,
   author = {Dydowiczová, Aneta and Brózman, Ondřej and Babica, Pavel and Sovadinová, Iva},
   article_location = {LONDON},
   article_number = {1},
   doi = {https://doi.org/10.1038/s41598-020-57536-3},
   keywords = {LIVER EPITHELIAL-CELLS; WB-F344 RAT-LIVER; FUNCTIONAL EXPRESSION; MOLECULAR PERMEABILITY; HA-RAS; CONNEXIN; INHIBITION; CHANNELS; CX43; IMAGE},
   language = {eng},
   issn = {2045-2322},
   journal = {Scientific reports},
   title = {Improved multiparametric scrape loading-dye transfer assay for a simultaneous high-throughput analysis of gap junctional intercellular communication, cell density and viability},
   url = {https://www.nature.com/articles/s41598-020-57536-3},
   volume = {10},
   year = {2020}
}

TY  - JOUR
ID  - 1729212
AU  - Dydowiczová, Aneta - Brózman, Ondřej - Babica, Pavel - Sovadinová, Iva
PY  - 2020
TI  - Improved multiparametric scrape loading-dye transfer assay for a simultaneous high-throughput analysis of gap junctional intercellular communication, cell density and viability
JF  - Scientific reports
VL  - 10
IS  - 1
SP  - 1-17
EP  - 1-17
PB  - NATURE PUBLISHING GROUP
SN  - 20452322
KW  - LIVER EPITHELIAL-CELLS
KW  - WB-F344 RAT-LIVER
KW  - FUNCTIONAL EXPRESSION
KW  - MOLECULAR PERMEABILITY
KW  - HA-RAS
KW  - CONNEXIN
KW  - INHIBITION
KW  - CHANNELS
KW  - CX43
KW  - IMAGE
UR  - https://www.nature.com/articles/s41598-020-57536-3
L2  - https://www.nature.com/articles/s41598-020-57536-3
N2  - Gap junctional intercellular communication (GJIC) is a vital cellular process required for maintenance of tissue homeostasis. In vitro assessment of GJIC represents valuable phenotypic endpoint that could be effectively utilized as an integral component in modern toxicity testing, drug screening or biomedical in vitro research. However, currently available methods for quantifying GJIC with higher-throughputs typically require specialized equipment, proprietary software and/or genetically engineered cell models. To overcome these limitations, we present here an innovative adaptation of traditional, fluorescence microscopy-based scrape loading-dye transfer (SL-DT) assay, which has been optimized to simultaneously evaluate GJIC, cell density and viability. This multiparametric method was demonstrated to be suitable for various multiwell microplate formats, which facilitates an automatized image acquisition. The assay workflow is further assisted by an open source-based software tools for batch image processing, analysis and evaluation of GJIC, cell density and viability. Our results suggest that this approach provides a simple, fast, versatile and cost effective way for in vitro high-throughput assessment of GJIC and other related phenotypic cellular events, which could be included into in vitro screening and assessment of pharmacologically and toxicologically relevant compounds.
ER  -

DYDOWICZOVÁ, Aneta; Ondřej BRÓZMAN; Pavel BABICA a Iva SOVADINOVÁ. Improved multiparametric scrape loading-dye transfer assay for a simultaneous high-throughput analysis of gap junctional intercellular communication, cell density and viability. \textit{Scientific reports}. LONDON: NATURE PUBLISHING GROUP, 2020, roč.~10, č.~1, s.~1-17. ISSN~2045-2322. Dostupné z: https://doi.org/10.1038/s41598-020-57536-3.

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